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Roles of the MSP-1₃₃ in the induction of anti-malaria response.

Tam, Hou Si. / 33 in title is subscript. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 174-187). / Abstracts in English and Chinese. / THESIS COMMITTEE --- p.i / ACKNOWLEDGEMENTS --- p.ii / ABSTRACT --- p.iii / 摘要 --- p.v / TABLE OF CONTENTS --- p.vii / LIST OF FIGURES --- p.xii / LIST OF TABLES --- p.xvii / LIST OF ABBREVIATIONS --- p.xviii / CHAPTER / Chapter 1. --- INTRODUCTION / Chapter 1.1 --- Malaria --- p.1 / Chapter 1.2 --- Malaria is a public health problem --- p.1 / Chapter 1.3 --- Malarial parasite --- p.3 / Chapter 1.4 --- Life cycle of P. falciparum --- p.3 / Chapter 1.4.1 --- The pre-erythrocytic stage --- p.3 / Chapter 1.4.2 --- The asexual erythrocytic stage --- p.3 / Chapter 1.4.3 --- The sexual transmission stage --- p.6 / Chapter 1.5 --- Chemoprophylaxis and chemotherapy of malaria --- p.7 / Chapter 1.6 --- Drug resistance of malaria parasite --- p.7 / Chapter 1.7 --- The progress for malaria vaccine --- p.10 / Chapter 1.8 --- Vaccine candidates for asexual erythrocytic stage --- p.11 / Chapter 1.9 --- Merozoite Surface Protein-1 (MSP-1) --- p.13 / Chapter 1.9.1 --- Structure of MSP-1 --- p.13 / Chapter 1.9.2 --- The processing of MSP-1 --- p.17 / Chapter 1.9.3 --- MSP-1 as a blood-stage vaccine --- p.19 / Chapter 1.9.4 --- The vaccine potency of MSP-133 --- p.23 / Chapter 1.10 --- Merits of E. coli expression system --- p.25 / Chapter 1.11 --- Aim of study --- p.26 / Chapter 2. --- MATERIALS AND METHODS / Chapter 2.1 --- Materials --- p.30 / Chapter 2.2 --- Methods --- p.39 / Chapter 3. --- EXPRESSION AND PURIFICATION OF RECOMBINANT MSP-l33kv+19 PROTEIN / Chapter 3.1 --- Introduction --- p.63 / Chapter 3.2 --- Results / Chapter 3.2.1 --- Construction of pET32a/MSP-l33kv+19 expression vector --- p.64 / Chapter 3.2.2 --- SDS-PAGE analysis of the expressed protein --- p.74 / Chapter 3.2.3 --- Western blot analysis of the expressed protein --- p.78 / Chapter 3.2.4 --- Modification of the expression conditions --- p.78 / Chapter 3.2.5 --- Protein purification by IMAC --- p.82 / Chapter 3.2.6 --- Cleavage of fusion partner from the rMSP-133kv+19 protein --- p.82 / Chapter 3.2.7 --- Verification of non-fused recombinant MSPl33kv+19 protein by N-terminal amino acid sequencing --- p.86 / Chapter 3.2.8 --- Separation of target protein from the fusion mixture by IMAC --- p.86 / Chapter 3.2.9 --- Separation of digestion product by Size Exclusion Chromatography --- p.89 / Chapter 3.2.10 --- Conformational test of the purified protein --- p.89 / Chapter 3.2.11 --- Separation of target protein from contaminants by Anion-Exchange Chromatography --- p.92 / Chapter 3.2.12 --- Separation of target protein from contaminants by Immuno-Affinity Chromatography --- p.95 / Chapter 3.3 --- Conclusion --- p.95 / Chapter 4. --- IMMUNOLOGICAL CHARACTERIZATION OF BACTERIAL EXPRESSED rMSP-l33kv+19 / Chapter 4.1 --- Introduction --- p.97 / Chapter 4.2 --- Results / Chapter 4.2.1 --- Immunogenicity of recombinant NfMSP-133kV+19 protein --- p.98 / Chapter 4.2.2 --- Specificity of anti-NfMSP-133kv+19 sera to MSP-l33kv. MSP-l33 and MSP-l19 --- p.98 / Chapter 4.2.3 --- Cross reactivity of anti-MSP-133kv+19 and anti-BVp42 serum --- p.103 / Chapter 4.2.4 --- Competitive ELISA --- p.103 / Chapter 4.2.5 --- Test for the presence of inhibitory B-cell epitopes on rMSP-l33kv+19 --- p.111 / Chapter 4.2.6 --- In vitro parasitic growth inhibition assay --- p.113 / Chapter 4.3 --- Conclusion --- p.115 / Chapter 5. --- EXPRESSION AND PURIFICATION OF RECOMBINANT MSP-l33kc+19 PROTEIN / Chapter 5.1 --- Introduction --- p.116 / Chapter 5.2 --- Results / Chapter 5.2.1 --- Construction of pET32a/MSP-133kv+19 expression vector --- p.117 / Chapter 5.2.2 --- Expression of recombinant MSP-133kc+19 protien (rMSP-133kc+19) --- p.124 / Chapter 5.2.3 --- Purification of rMSP-l33kc+19 by IMAC --- p.127 / Chapter 5.2.4 --- Cleavage of fusion partner from target protein --- p.127 / Chapter 5.2.5 --- Construction of pRSETA/MSP-l3X33kc+19 expression vector --- p.135 / Chapter 5.2.6 --- SDS-PAGE analysis of the protein expression --- p.146 / Chapter 5.3 --- Conclusion --- p.153 / Chapter 6. --- DISCUSSION / Chapter 6.1 --- Expression of rMSP-l33kv+19 --- p.154 / Chapter 6.2 --- Purification of rMSP-l3.3kv+19 --- p.156 / Chapter 6.3 --- Conformational test of rMSP-133kv+19 --- p.157 / Chapter 6.4 --- Biological and immunological activity of NfMSP-133kv+19 --- p.158 / Chapter 6.5 --- Expression of rMSP-133kc+19 --- p.166 / Chapter 6.6 --- Future prospects --- p.167 / REFERENCES --- p.174 / APPENDICES / Chapter 1. --- HiTrap NHS-activated HP for ligand coupling procedure --- p.188 / Chapter 2. --- Reuse of Ni+-NTA Resin procedure --- p.190 / Chapter 3. --- Sequence alignment of MSP-133 (MAD20 & Welcome/Kl alleles) --- p.191 / Chapter 4. --- Nucleotide sequence and amino acid sequence of P. falciparum MSP-l33kv+19 --- p.192 / Chapter 5. --- Nucleotide sequence and amino acid sequence of P. falciparum MSP-l33kc+19 --- p.193 / Chapter 6. --- "Nucleotide sequence and amino acid sequence of P. falciparum MSP-142 (3D7 isolate, MAD20 allele)" --- p.194 / Chapter 7. --- Amino acid sequence of Plasmodium falciparum MSP-l42 --- p.195

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_326094
Date January 2007
ContributorsTam, Hou Si., Chinese University of Hong Kong Graduate School. Division of Biochemistry.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xx, 195 leaves : ill., maps ; 30 cm.
RightsUse of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/)

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